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通过逆转录-聚合酶链反应检测新鲜人结肠癌组织中的胃泌素mRNA

Detection of gastrin mRNA in fresh human colonic carcinomas by reverse transcription-polymerase chain reaction.

作者信息

Monges G, Biagini P, Cantaloube J F, Chicheportiche C, Frances V, Brandini D, Parc P, Seitz J F, Giovannini M, Sauvan R

机构信息

Paoli-Calmettes Institute, Marseille, France.

出版信息

J Mol Endocrinol. 1993 Oct;11(2):223-9. doi: 10.1677/jme.0.0110223.

Abstract

To investigate the hypothesis that gastrin might be synthesized by tumour tissues in cancer of the colon, samples from six human colon tumours, one hepatic metastasis, four normal colonic mucosal samples and two antral and one fundic gastric mucosal samples from nine patients were analysed to determine whether gastrin mRNA was present. RNA was extracted from surgical specimens by ultracentrifugation on a CsCl cushion, purified using the guanidinium thiocyanate method, reverse-transcribed and amplified by polymerase chain reaction. Gastrin mRNA was detected in each colonic carcinoma sample (including the hepatic metastasis), while no such signal was observed in normal colon biopsies. Positive and negative controls (gastric antrum and fundus respectively) gave the expected results. In each of the positive samples, the chemiluminescent revelation of amplified products after Southern blotting corresponded to gastrin mRNA without the intron. These findings demonstrate the ability of primary and metastatic human colonic tumours to produce gastrin mRNA. Since malignant cell lines have been reported to produce gastrin peptide, and since gastrin receptors were present in some cases, our results support the idea that gastrin may be involved in an autocrine mechanism.

摘要

为了研究胃泌素可能由结肠肿瘤组织合成这一假说,对来自9名患者的6份人类结肠肿瘤样本、1份肝转移样本、4份正常结肠黏膜样本以及2份胃窦和1份胃底胃黏膜样本进行了分析,以确定是否存在胃泌素mRNA。通过在氯化铯垫层上超速离心从手术标本中提取RNA,采用硫氰酸胍法进行纯化,逆转录并通过聚合酶链反应进行扩增。在每个结肠癌样本(包括肝转移样本)中均检测到胃泌素mRNA,而在正常结肠活检样本中未观察到此类信号。阳性和阴性对照(分别为胃窦和胃底)给出了预期结果。在每个阳性样本中,Southern印迹后扩增产物的化学发光显示对应于不含内含子的胃泌素mRNA。这些发现证明了原发性和转移性人类结肠肿瘤产生胃泌素mRNA的能力。由于已有报道称恶性细胞系可产生胃泌素肽,且在某些情况下存在胃泌素受体,我们的结果支持胃泌素可能参与自分泌机制这一观点。

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